Device and method for detecting redox reactions in solution

ABSTRACT

Described herein is a device comprising a plurality of first reaction electrodes arranged in an array, the plurality of first reaction electrodes configured to be exposed to a solution and having a capacitance; first circuitry configured to controllably connect the plurality of first reaction electrodes to a bias source and controllably disconnect the plurality of first reaction electrodes from the bias source; and second circuitry configured to measure a rate of charging or discharging of the capacitance. Also described herein is a method of using this device to sequence DNA.

CROSS-REFERENCE TO RELATED APPLICATIONS

The present application is a divisional application of U.S. patentapplication Ser. No. 14/267,327, filed May 1, 2014, which is adivisional application of U.S. patent application Ser. No. 13/628,728,filed Sep. 27, 2012, now granted U.S. Pat. No. 8,741,117, issued Jun. 3,2014, which Applicant claims the benefit of priority and both of whichare incorporated herein by reference in their entireties.

Reference is made to commonly owned and co-pending U.S. application Ser.No. 12/655,578 titled “Nanogap Chemical and Biochemical Sensors,” filedDec. 31, 2009, now pending; U.S. patent application Ser. No. 11/226,696,titled “Sensor Arrays and Nucleic Acid Sequencing Applications,” filedSep. 13, 2005, now pending; which is a continuation-in-part applicationthat claims the benefit of U.S. patent application Ser. No. 11/073,160,titled “Sensor Arrays and Nucleic Acid Sequencing Applications,” filedMar. 4, 2005; U.S. patent application Ser. No. 11/967,600, titled“Electronic Sensing for Nucleic Acid Sequencing,” filed Dec. 31, 2007now pending; U.S. patent application Ser. No. 12/319,168, titled“Nucleic Acid Sequencing and Electronic Detection,” filed Dec. 31, 2008,now pending; U.S. patent application Ser. No. 12/459,309, titled“Chemically Induced Optical Signals and DNA Sequencing,” filed Jun. 30,2009, now pending; U.S. patent application Ser. No. 12/655,459, titled“Solid-Phase Chelators and Electronic Biosensors,” filed Dec. 30, 2009,now pending; U.S. patent application Ser. No. 12/823,995, titled“Nucleotides and Oligonucleotides for Nucleic Acid Sequencing,” filedJun. 25, 2010, now pending; U.S. patent application Ser. No. 12/860,462,titled “Nucleic Acid Sequencing,” filed Aug. 20, 2010, now pending; thedisclosures of which are incorporated herein by reference in theirentirety. Appropriate components for device/system/method/processaspects of the each of the foregoing U.S. patents and patentpublications may be selected for the present disclosure in embodimentsthereof.

BACKGROUND

DNA sequencing is the process of reading the nucleotide bases in a DNAmolecule. It includes any method or technology that is used to determinethe order of the four bases—adenine, guanine, cytosine, and thymine—in astrand of DNA.

Knowledge of DNA sequences is useful for basic biological research, andin numerous applied fields such as diagnostic, biotechnology, forensicbiology, and biological systematics. The advent of DNA sequencing hasaccelerated biological research and discovery. The rapid speed ofsequencing attained with modern DNA sequencing technology has beeninstrumental in the sequencing of the human genome, in the Human GenomeProject. Related projects, often by scientific collaboration acrosscontinents, have generated the complete DNA sequences of many animal,plant, and microbial genomes.

New development in the medical field (e.g., personalized medicine) andbasic biological research (e.g., animal or plant genome projects) callsfor rapid sequencing of large number (e.g., above 10,000) of DNAfragments in a practical period of time (e.g., several hours to severaldays), which is usually referred to as a high-throughput sequencing.Traditional chemistry-based and optic-based DNA sequencing methods suchas the Maxam-Gilbert method and Chain-termination methods suffer fromtheir requirements of complex sample preparation and slow rate of basedetection, and are generally unsuitable in these applications.

Exemplar high-throughput sequencing techniques include MassivelyParallel Signature Sequencing (MPSS) developed by Lynx Therapeutics,Polony sequencing developed by Prof. George Church at HarvardUniversity, parallelized pyrosequencing developed by 454 Life Sciences(now Roche Diagnostics), SOLiD sequencing developed by AppliedBiosystems (now Life Technologies), pH-based semiconductor sequencingdeveloped by Ion Torrent (now Life Technologies), nanopore sequencing,etc.

SUMMARY

Described herein is a device comprising: a plurality of first reactionelectrodes arranged in an array, the plurality of first reactionelectrodes configured to be exposed to a solution and having acapacitance; first circuitry configured to controllably connect theplurality of first reaction electrodes to a bias source and controllablydisconnect the plurality of first reaction electrodes from the biassource; and second circuitry configured to measure a rate of charging ordischarging of the capacitance of the plurality of first reactionelectrodes.

Also described herein is a method comprising contacting an electrode toa solution comprising one or more analyte; connecting the electrode to abias source such that a voltage of the electrode is substantially at avoltage of the bias source; disconnecting the electrode from the biassource; measuring change of the voltage of the electrode.

Further described herein is a method comprising contacting a pluralityof electrodes to a solution, each of the plurality of electrodes havinga DNA molecule attached thereto or in proximity thereof, wherein DNAmolecule has a primer hybridized thereto; adding at least one type ofnucleoside triphosphate to the solution; setting the solution to atemperature suitable for the at least one type of nucleosidetriphosphate to be incorporated into at least some of the primers;measuring a rate of charge or discharge of capacitance of each of theplurality of electrodes. Incorporating the at least one type ofnucleoside triphosphate can release a reaction product that undergoes aredox reaction at the electrodes. The redox reaction may cause chargingor discharging of the capacitance. The nucleoside triphosphate may be inits natural form or modified with a moiety which is either directlyredox-active upon release or can be subsequently activated.

BRIEF DESCRIPTION OF FIGURES

FIG. 1 shows an exemplary three-electrode electrochemistry cell.

FIG. 2A shows an exemplary three-electrode electrochemistry cellaccording to an embodiment, wherein electrical current through one ofthe electrodes is measured by measuring the rate of charging ordischarging of capacitance of that electrode.

FIG. 2B shows an exemplary graph of the voltage of one of the threeelectrodes as a function of time, when the redox reaction discharges thecapacitance of that electrode.

FIG. 3A shows an array of the circuitry in the dotted box 300 in FIG.2A.

FIG. 3B shows that circuitry to measure the rate of charge or dischargeof one of the electrodes may be shared in the array of FIG. 3A.

FIG. 3C shows an array of the circuitry in the dotted box 300 in FIG.2A, except that a pixel of the array has at least two electrodes, therate of charging or discharging of capacitance of which are measured.

FIG. 3D shows an array of the circuitry in the dotted box 300 in FIG.2A, except that a pixel of the array has another biased electrodeintegrated in the pixel.

FIG. 4A shows an exemplary embodiment of the array.

FIG. 4B shows an exemplary schematic of a pixel in the array of FIG. 4A.

FIG. 5 shows a flowchart of a method of operating the array of FIG. 4A.

FIG. 6 shows an exemplary scheme to sequence DNA with the array of FIG.4A.

DETAILED DESCRIPTION

A DNA molecule may be sequenced by detecting reaction products fromincorporation of individual nucleotides into a polynucleotide chaincomplementary to the DNA molecule or cleavage of individual nucleotidesfrom the DNA molecule. Various enzymes (e.g., polymerase,deoxyribonuclease) may be used to facilitate the incorporation orcleavage.

One method to detect these reaction products is electrochemistry.Electrochemistry is a branch of chemistry that studies chemicalreactions which take place in a solution at the interface of an electronconductor (a metal or a semiconductor) and an ionic conductor (theelectrolyte), and which involve electron transfer between the electrodeand the electrolyte or species in solution. When the reaction productsundergo redox reactions in an electrochemistry cell, electricalsignatures (e.g., cyclic voltammogram) of the reaction products may beused to identify them. For example, the reaction products may be thosereleased from incorporation of chemically-modified nucleotides into DNA,as described in U.S. patent application Ser. No. 11/967,600, titled“Electronic Sensing for Nucleic Acid Sequencing,” which is herebyincorporated by reference in its entirety.

FIG. 1 shows an exemplary three-electrode electrochemistry cell: areference electrode 110, a second reaction electrode 120 and a firstreaction electrode 130 in a solution (electrolyte) 150. Theelectrochemistry cell can be used to conduct various electrochemicalreactions. One kind electrochemical reaction is called voltammetry. Involtammetry, information about an analyte can be obtained by measuringthe current through the first reaction electrode 130 and/or the currentthrough the second reaction electrode 120. The reference electrode 110is a half cell with a fixed reduction potential to the solution. Thereference electrode 110 may act as reference in measuring andcontrolling the potential on the second reaction electrode 120. Thereference electrode 110 is usually connected to electrical groundthrough a high impedance (e.g., an unity gain op amp 111). Electricalcurrent through the reference electrode 110 is negligible. The referenceelectrode 110 suitable in a aqueous electrolyte may be, for example,standard hydrogen electrode (SHE), normal hydrogen electrode (NHE),reversible hydrogen electrode (RHE), saturated calomel electrode (SCE),copper-copper(II) sulfate electrode (CSE), silver-silver chloride(Ag/AgCI) electrode, etc. The reference electrode 110 may also be apseudoreference or quasireference electrode (QRE) which is less stableand less reproducible in the electrode-solution potential but sufficientfor the measurement. The second reaction electrode 120 is electricallybiased relative to the reference electrode 110. The second reactionelectrode 120 may be biased using any suitable electrical circuitry 121.The bias on the second reaction electrode 120 may be used to control thevoltage of the solution 150 relative to the voltage of the firstreaction electrode 130. In this case the second electrode 120 is calledan auxiliary or counter electrode and controls the potential of thesolution. In some situations, the function of electrodes 110 and 120 canbe combined into a single electrode. The electrical bias on the secondreaction electrode 120 may be fixed or varied during electrochemicalreactions. The second reaction electrode 120 is an electricallyconducting material such as metal or semiconductor. For instance, thesecond reaction electrode 120 may comprise platinum, gold, dopeddiamond, glassy carbon, indium tin oxide, or a combination thereof.Furthermore, the second reaction electrode 120 may be coated with anorganic or inorganic film to improve its electrochemical properties. Thesecond reaction electrode 120 may be inert (i.e., not reactive with theelectrolyte or analyte) or reactive. The first reaction electrode 130passes electrical current needed to balance the current observed at thesecond reaction electrode 120. The electrical current through the firstreaction electrode 130 may be measured using any suitable electricalcircuitry 131. The first reaction electrode 130 can be any electricallyconducting material but preferably is not dissolved into the electrolyteduring electrochemical reactions. For instance, the first reactionelectrode 130 may comprise platinum, gold, doped diamond, glassy carbon,indium tin oxide, or a combination thereof. Furthermore, the firstreaction electrode 130 may be coated with an organic or inorganic filmto improve its electrochemical properties.

The second reaction electrode 120 need not be used as a traditionalcounter electrode (with potential set by electronic feedback to controlthe solution voltage). A voltage applied to the second reactionelectrode 120 may be used to initiate redox reaction cycles between thesecond reaction electrode 120 and the first reaction electrode 130.Preferably, electrodes 120 and 130 are in very close proximity, e.g.with 50 nm from each other. Namely, a chemical species receives one ormore electrons (i.e., reduced) at the second reaction electrode 120,diffuses to the first reaction electrode 130, loses one or moreelectrons (i.e., oxidized) at the first reaction electrode 130 anddiffuses back to the second reaction electrode 120; or a chemicalspecies receives one or more electrons (i.e., reduced) at the firstreaction electrode 130, diffuses to the second reaction electrode 120,loses one or more electrons (i.e., oxidized) at the second reactionelectrode 120 and diffuses back to the first reaction electrode 130. Theredox cycle may be used to increase the current through the firstreaction electrode 130 and/or the second reaction electrode 120 tofacilitate measurement of this current.

The electrochemistry cell may have more than three electrodes. Theelectrical current through each electrode may be measured independently.The electrical current on each electrode yields information about redoxreaction in the vicinity of that electrode.

In certain situations, for example, high reaction potentials of theanalyte, unfavorable voltage on the working electrode, low concentrationof the analyte, etc. may cause the electrical current though one of theelectrodes to be very small. Small electrical current may be difficultto measure reliably, especially in a solution environment, where noisetends to be high. Small electrical currents on spatially proximateelectrodes may be especially challenging to measure.

According to an embodiment shown in FIG. 2A, electrical current throughan electrode may be measured by measuring the rate of charging ordischarging of capacitance of that electrode. For example, electricalcurrent through the first reaction electrode 130 may be measured bymeasuring the rate of charging or discharging of capacitance 233 of thefirst reaction electrode 130. Although the capacitance 233 of the firstreaction electrode 130 is depicted in FIG. 2A as a capacitor separatefrom the capacitance, it need not comprise a physical capacitorcomponent, but a combination of self-capacitance of the first reactionelectrode 130 and capacitance of the interface between the firstreaction electrode 130 and the solution 150.

The circuitry connected to the first reaction electrode 130 as depictedin FIG. 2A is one example that can be used to measuring the rate ofcharging or discharging of the capacitance 233. In an embodiment, switch232 is closed to connect the first reaction electrode 130 to a biassource 231. At this state, the voltage on the first reaction electrode130 is at the voltage of the bias source 231, denoted as V₀. The switch232 may be any circuitry that can electrically connect and disconnectthe first reaction electrode 130 to the bias source 231. For example,the switch 232 may be a toggle switch, a relay or a transistor. Afterthe switch 232 is opened to disconnect the first reaction electrode 130from the bias source 231, redox reactions (electron transfer between theelectrode and a chemical species in the solution) occurring at the firstreaction electrode 130 start to charge or discharge the capacitance 233and as a result the voltage of the first reaction electrode 130 deviatesfrom V₀. The rate of charging and discharging of the capacitance 223 canbe derived from the change of the voltage of the first reactionelectrode 130. The voltage of the first reaction electrode 130 may bemeasured using any suitable circuitry 234. Circuitry 234 is not limitedto a voltmeter. In an embodiment, the circuitry 234 may comprise A/Dconverter. In an embodiment, the circuitry 234 may comprise a buffer.The buffer may drive an A/D converter shared with other electrodes. FIG.2B shows an exemplary graph of the voltage of the first reactionelectrode 130 as a function of time, when the redox reaction dischargesthe capacitance 233. The more molecules undergo redox reactions in thevicinity of the first reaction electrode 130, the greater the rate ofcharging or discharging of the capacitance 233. For example, trace 291in FIG. 2B shows discharge of the capacitance 233 when essentially nomolecules undergo redox reactions in the vicinity of the first reactionelectrode 130. The slight drop of the voltage on the first reactionelectrode 130 is due to leakage current through the solution 150 and/orleakage current from the circuitry 231 and/or 234. Traces 292 and 293 inFIG. 2B show discharge of the capacitance 233 when some moleculesundergo redox reactions in the vicinity of the first reaction electrode130. Trace 293 shows a greater rate of discharge that trace 292 becausemore molecules undergo redox reactions in the vicinity of the firstreaction electrode 130 when trace 293 is recorded than when trace 292 isrecorded. The rate of discharge may be measured directly (e.g., theslope of the traces in FIG. 2B) or by any other suitable techniques. Forexample, the rate of discharge may be measured by the amount of voltagechange from V₀ at a time point t_(int) after the first reactionelectrode 130 disconnects from the bias source 231. In alternative, toreduce drift or noise in circuitry 234, switch 232 can be closed at atime just after t_(int), reseting the voltage of the first reactionelectrode 130 to V₀, and the rate of discharge may be measured by thedifference between the voltage at t_(int) and the voltage after thereset. In another alternative, the rate of discharge may be measured byby the amount of time for the voltage of the first reaction electrode130 to change from V₀ by a predetermined amount to a voltage V_(th)after the first reaction electrode 130 disconnects from the bias source231.

In an embodiment, the circuitry in the dotted box 300 in FIG. 2A,including the first reaction electrode 130 may be integrated in amicrochip. In an embodiment, such a microchip may have a plurality ofcopies of the circuitry in the dotted box 300. These copies may bearranged in an array (e.g., a rectangular or hexagonal grid) such as thearray shown in FIG. 3A. Preferably, the bias source 231 is shared amongthese copies. For the purpose of this disclosure, a copy may be referredto as a “pixel.” In an embodiment as shown in FIG. 3A, a pixel may haveits own circuitry 234 (i.e., not shared with other pixels) integrated inthe pixel. The measurements of the circuitry 234 in each may betransmitted to a bus through a multiplexer. In an embodiment depicted inFIG. 3B, the circuitry 234 may be shared among these copies through amultiplexer 390. A multiplexer (or MUX) is a device that selects one ofseveral analog or digital input signals and forwards the selected inputinto a single line. In an embodiment, the circuitry 234 includes ahigh-impedance buffer configured to measure the voltage of the firstreaction electrode 130 without discharging the capacitance 233. In anembodiment, the high-impedance buffer is included in each pixel butother portions (such as A/D converter) of the circuitry 234 are sharedamong many pixels through a multiplexer. In an embodiment, a pixelcomprises a memory configured to store the measurement by the circuitry234 of the first reaction electrode 130. For example, the memory may beconfigured to measure and store the value of the voltage of the firstreaction electrode 130 at t_(int) or the time at which the voltage ofthe first reaction electrode 130 crossed a predetermined thresholdvalue.

In an embodiment schematically shown in FIG. 3C, a pixel may have atleast two first reaction electrodes 130 a and 130 b. The first reactionelectrodes 130 a and 130 b are arranged to be in vicinity of each other.As used in this disclosure, the word “vicinity” means distance nogreater than the diffusion length of the analyte at t_(int). Correlationof the voltages on the first reaction electrodes 130 a and 130 b may beused to determine the validity of data collected from this pixel. Forexample, if the voltages on the first reaction electrodes 130 a and 130b differ by a threshold value, data collected from this pixel isconsidered invalid and discarded.

In an embodiment schematically shown in FIG. 3D, a pixel may have thesecond reaction electrode 120 integrated in the pixel. In an embodiment,the second reaction electrode 120 can be shared among more than onepixel.

In an embodiment, a pixel has an area from 0.01 to 100 μm², such asabout 1 μm². In one embodiment, the array has more than 100 pixels, 10⁶pixels, or 10¹⁰ pixels. In an embodiment, the pixels do not comprise aphotodiode.

FIG. 4A shows an exemplary embodiment of the array. The pixels may beinterrogated by enabling a row of pixels using row decoder 430 whichconnects the pixels to a shared A/D converter by enabling a switch(e.g., a transistor 290 in FIG. 4B). 420 may be an array of A/Dconverters (one for each column of pixels) or a multiplexer thatconnects the column outputs to one or more shared A/D converters. 420and 430 are under control of a controller 410. FIG. 4B shows anexemplary schematic of a pixel 400 in the array of FIG. 4A. Themultiplexing functionality may be provided by transistor 290. Transistor280 may act as a voltage buffer to read the voltage at electrode 130without discharging the capacitance of electrode 130. Switch 232 may beimplemented as a transistor. The functionality provided by switch 232,transistors 280 and 290 can be implemented using different electronicdevices than shown as examples in FIG. 4B. One row of pixels in thearray may be interrogated at a time, and the entire array may beinterrogated by interrogating each row of pixels. Furthermore, theentire array can be interrogated multiple times to obtain theinformation such as that contained in FIG. 2B.

In an embodiment, as schematically shown in the flowchart of FIG. 5, amethod of operating the array comprises: contacting the first reactionelectrode 130 to the solution 150 comprising one or more analyte;connecting the first reaction electrode 130 to the bias source 231, suchthat the voltage of the first reaction electrode 130 is substantially atthe voltage V₀ of the bias source 231; disconnecting the first reactionelectrode 130 from the bias source 231; measuring change of the voltageof the first reaction electrode 130, wherein the change of the voltageis produced from charging or discharging of capacitance of the firstreaction electrode 130 due to redox reactions in the vicinity of thefirst reaction electrode 130. The method may further comprise storingthe change of the voltage of the first reaction electrode 130 in amemory.

An exemplary method of using the array to sequence DNA may comprise thefollowing. A pixel of the array may have one or more copies of a DNAmolecule to be sequenced attached to or in proximity of the firstreaction electrodes 130 therein, or the second reaction electrode 120 ifthe pixel has its own second reaction electrode 120 (i.e., the secondreaction electrode 120 not shared with other pixels), the DNA moleculeto be sequenced having a primer hybridized thereto. The pixel preferablydoes not have DNA molecules of more than one sequence. The array is incontact with the solution 150. The solution 150 comprises reagents suchas polymerase, and salt (e.g., MgCl₂). Only one type of nucleosidetriphosphate (i.e., one type among deoxyadenosine triphosphate (dATP),deoxyguanosine triphosphate (dGTP), deoxythymidine triphosphate (dTTP)and deoxycytidine triphosphate (dCTP)) is added to the solution 150. Thenucleoside triphosphate may be in its natural form or modified with amoiety that can undergo a redox reaction in the solution 150. Thesolution 150 is set to a temperature suitable for the polymerase toincorporate the nucleoside triphosphate into the one or more DNAprimers. Only pixels with DNA molecules whose next unpaired base iscomplementary to the added nucleoside triphosphate will have thenucleoside triphosphate incorporated to the primers. For example, asillustrated in FIG. 6, in which dATP is added into solution 150 as thenucleoside triphosphate, four pixels each with a first reactionelectrode (130A, 130T, 130G and 130C) is shown. On first reactionelectrodes 130A, 130T, 130G and 130C DNA molecules 6A, 6T, 6G and 6C arerespectively attached. Each of these DNA molecules has a primer beingextended by the polymerase. On DNA molecules 6A, 6T, 6G and 6C, the nextunpaired bases are A, T, G and C, respectively. The dATP in the solution150 will thus only be incorporated to the primer on the DNA molecule 6A.The polymerase splits the terminal phosphate groups (with or without themoiety) from the incorporated dATP. The terminal phosphate groups splitfrom nucleoside triphosphate is often referred to as phosphate residue.The phosphate residue and/or the moiety undergoes redox reaction in thevicinity of the first reaction electrode 130T. As a result, capacitance233T of the first reaction electrode 130T charges or discharges at arate beyond that caused by leakage current (e.g., trace 291 in FIG. 2B).Since no incorporation of dATP to the primers on the other three firstreaction electrodes 130A, 130G and 130C, capacitance 233A, 233G and 233Cof these first reaction electrodes 130A, 130G and 130C do not charges ordischarges at a rate beyond that caused by leakage current. Thedifference between the rate of charge of discharge on the first reactionelectrodes 130A, 130T, 130G and 130C indicates that the next unpairedbase on DNA 6T is thymine. Solution 150 is removed and the array iswashed to remove any residue of solution 150. These steps are repeatedwith the other three nucleoside triphosphates to complete sequencing ofall the DNA molecules on the pixels of the array. Alternatively, morethan one type of nucleoside triphosphate modified with differentmoieties may be added to the solution 150. The electrodes 130A, 130T,130G and 130C may detect that the next unpaired base thereon bydetecting the rate of charge of discharge on these electrodesrespectively, caused by redox reactions of the different moieties on thenucleoside triphosphate. Other DNA sequencing methods that can beapplied to the array include those disclosed in U.S. patent applicationSer. No. 11/967,600, titled “Electronic Sensing for Nucleic AcidSequencing,” filed Dec. 31, 2007, the disclosures of which areincorporated herein by reference in its entirety.

While various aspects and embodiments have been disclosed herein, otheraspects and embodiments will be apparent to those skilled in the art.The various aspects and embodiments disclosed herein are for purposes ofillustration and are not intended to be limiting, with the true scopeand spirit being indicated by the following claims.

We claim:
 1. A method comprising: contacting an electrode of a devicecomprising a first circuitry and a second circuitry to a solution, theelectrode has a capacitance and is in an array comprising a plurality ofelectrodes, each of the plurality of electrodes having a DNA moleculeattached thereto or in proximity thereof, wherein DNA molecule has aprimer hybridized thereto; adding at least one type of nucleosidetriphosphate to the solution; setting the solution to a temperaturesuitable for the one type of nucleoside triphosphate to be incorporatedinto at least some of the primers; measuring a rate of charge ordischarge of capacitance of each of the plurality of electrodes.
 2. Themethod of claim 2, further comprising removing the solution and washingthe plurality of electrodes.
 3. The method of claim 1, wherein the firstcircuitry is configured to (a) set a DC voltage on the electrode, andconfigured to charge or discharge the capacitance through a redoxreaction occurring at the electrode; and the second circuitry isconfigured to measure a rate of charging or discharging of thecapacitance of the electrode, or (b) controllably connect the electrodeto the bias source and controllably disconnect the electrode from thebias source; and the second circuitry is configured to measure the rateof charging or discharging of the capacitance of the electrode.
 4. Themethod of claim 1, further comprising storing the change of the voltageof the electrode in a memory.
 5. The method of claim 1, furthercomprising repeating steps of contacting, connecting, disconnecting andmeasuring on at least 100 electrodes.
 6. The method of claim 1, whereinthe second circuitry is configured to measure the rate of charging ordischarging of the capacitance by measuring a voltage of the electrodeas a function of time.
 7. The method of claim 1, wherein the secondcircuitry is configured to measure the rate of charging or dischargingof the capacitance by measuring an amount of voltage change from avoltage of the bias source at a time point.
 8. The method of claim 1,wherein the second circuitry is configured to measure the rate ofcharging or discharging of the capacitance by measuring an amount oftime for a voltage of the electrode to change from a voltage of the biassource by a predetermined amount.
 9. The method of claim 1, wherein thedevice is a microchip.
 10. The method of claim 1, wherein thecapacitance includes self-capacitance of the electrode and capacitanceof interfaces between the electrode and the solution.
 11. The method ofclaim 1, wherein a geometric area of the electrode is between 0.01 μm²and 100 μm².
 12. The method of claim 1, further comprising a referenceelectrode with a fixed reduction potential to the solution.
 13. Themethod of claim 12, wherein the reference electrode is selected from thegroup consisting of standard hydrogen electrode (SHE), normal hydrogenelectrode (NHE), reversible hydrogen electrode (RHE), saturated calomelelectrode (SCE), copper-copper(II) sulfate electrode (CSE),silver-silver chloride (Ag/AgCl) electrode, silver pseudoreferenceelectrode, and quasi-reference (QRE).
 14. The method of claim 1, whereinthe electrode are a material that is not consumed by the solution. 15.The method of claim 1, wherein the first circuitry comprises a switch.16. The method of claim 15, wherein the second circuitry is configuredto measure the voltage of the electrode without discharging thecapacitance.
 17. The method of claim 1, wherein the second circuitrycomprises a memory.
 18. The method of claim 1, wherein the secondcircuitry is shared among more than one electrodes.
 19. The method ofclaim 1, wherein the device further comprises an additional electrodeconfigured to be electrically biased relative to the solution.
 20. Themethod of claim 19, wherein the additional electrode is not consumed.21. The method of claim 19, wherein the electrode and/or the additionalelectrode are coated with an organic or inorganic coating.
 22. Themethod of claim 19, wherein the electrode is selected from the groupconsisting of platinum, gold, indium tin oxide, diamond-like carbondoped with impurities, glassy carbon, silver, carbon nanotube, graphene,and conducting polymers; and/or wherein the additional electrode isselected from the group consisting of platinum, gold, indium tin oxide,diamond-like carbon doped with impurities, glassy carbon, silver, carbonnanotube, graphene, and conducting polymers.